The DNAJC25 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human NCI-H1299 non-small cell lung cancer (NSCLC) cell line, featuring targeted disruption of the DNAJC25 gene. This polyclonal format provides a heterogeneous pool of knockout cells, enabling studies that require broad representation of loss-of-function phenotypes without clonal selection bias.
The NCI-H1299 line is a well-established model of NSCLC, derived from lymph node metastasis and characterized by p53 deficiency. These epithelial cells are widely used for investigating tumorigenesis, metastasis, and drug responses, and their robust growth supports both in vitro and xenograft studies.
DNAJC25 encodes a co-chaperone that directly interacts with Hsp70 proteins, including HSPA1A and HSPA8, to regulate protein folding, degradation, and stress adaptation. Its activity is controlled by stress-responsive transcription factors HSF1, XBP1, and ATF6, integrating it into the UPR and ER stress pathways. DNAJC25 also associates with co-chaperones BAG3 and STIP1 and modulates downstream effectors IRE1 and BCL2 family members, thereby influencing both proteostasis and apoptosis. Disruption of DNAJC25 impairs Hsp70-mediated client handling and may enhance stress-induced apoptosis.
In NCI-H1299 lung cancer cells, loss of DNAJC25 is pertinent to understanding how NSCLC cells cope with proteotoxic stress and evade apoptosis. Tumor cells often upregulate the Hsp70 system to maintain proteostasis; knockout of DNAJC25 may compromise this adaptation, revealing therapeutic vulnerabilities. This model allows dissection of DNAJC25??s role in stress resilience, ER stress signaling, and apoptotic regulation within a p53-deficient context, providing insight into co-chaperone dysfunction in lung cancer.
This knockout product supports diverse applications, including proteomic identification of DNAJC25-dependent Hsp70 client proteins, analysis of UPR and ER stress pathway activation, and functional studies of apoptosis networks. Standard assays such as viability measurements (MTT, CellTiter-Glo), Annexin V apoptosis detection, co-immunoprecipitation, RT-qPCR, and Western blotting are readily applicable. The cells also serve as a platform for drug sensitivity screening to identify compounds that exploit DNAJC25 loss. For further information, please contact Ascent Research.
